Proposal for an Administrative Supplement for Equipment Purchases for NIGMS Awardees NOT-GM-22-017

家长资助项目摘要R 01 GM 132329 膜蛋白在生物体中是必不可少的。蛋白质-脂质相互作用对膜的形成至关重要 蛋白质结构和功能的完整性。高分辨率的膜蛋白结构是在高 需要了解它们的生物学和基于结构的药物开发。提取 生物化学和生物物理学研究经常需要来自细胞膜的膜蛋白。 目前，洗涤剂是最常用的提取膜蛋白的方法。洗涤剂 有几个缺点，包括脂质双层的破坏和必需脂质的去除 膜蛋白分子。我们正在制造一种不含洗涤剂的天然细胞膜 纳米颗粒技术用于膜蛋白研究。天然细胞膜纳米颗粒 系统由三部分组成：1）具有膜活性的聚合物。2)创建本机 细胞膜纳米颗粒。3)天然细胞膜的Cryo-EM单粒子研究 纳米粒子我们建议建立一个适用于细菌和真核细胞膜的系统 proteins.具体目标1侧重于构建技术，以创建天然细胞膜纳米颗粒 利用我们选择的模型膜蛋白和构建膜活性聚合物库。我们 我们已经创建了四个原型膜活性聚合物;我们打算扩大活性膜 使用市售苯乙烯-马来酸酐共聚物和由其制备的聚合物的聚合物 在实验室中使用类似的程序。具体的Ami 2侧重于创建程序， 所选细菌模型膜蛋白的高分辨率结构。第三个具体目标是 创建选定的真核模型膜蛋白的高分辨率结构的程序。的 提出的研究是有趣的，因为它提供了一种新的无洗涤剂的方法来研究膜 蛋白质在其天然脂质环境中。正如我们的第一个数据显示，蛋白质-脂质的结构知识 膜蛋白的相互作用对于理解膜的结构和功能至关重要。 proteins.这种方法有可能彻底改变膜蛋白研究领域， 对基于膜蛋白结构的药物发现和开发具有重大影响。

Project Summary for Parent Grant R01GM132329 Membrane proteins are essential in living organisms. Protein-lipid interactions are critical for membrane protein structural and functional integrity. High-resolution membrane protein structures are in high demand for understanding their biology and drug development based on structure. Extraction of membrane proteins from cell membranes is frequently required for biochemical and biophysical research. Currently, detergents are the most often used method for extracting membrane proteins. Detergents have several downsides, including the destruction of lipid bilayers and the removal of essential lipid molecules from membrane proteins. We are creating a detergent-free native cell membrane nanoparticles technology for membrane protein research. The native cell membrane nanoparticles system is made up of three parts: 1) Polymers that are membrane-active. 2) Methods for creating native cell membrane nanoparticles. 3) Cryo-EM single particle investigation of native cell membrane nanoparticles. We suggest creating a system that is applicable to both bacterial and eukaryotic membrane proteins. Specific Aim 1 focuses on building techniques to create native cell membrane nanoparticles utilizing our chosen model membrane proteins and constructing a membrane-active polymer library. We have created four prototype membrane active polymers; we intend to expand the active membrane polymers utilizing both commercially available styrene-maleic anhydride copolymers and polymers made in laboratories using similar procedures. Specific Ami 2 focuses on creating procedures for determining the high-resolution structure of selected bacterial model membrane proteins. The third specific aim is to create procedures for high-resolution structures of selected eukaryotic model membrane proteins. The proposed study is interesting because it offers a novel detergent-free method for investigating membrane proteins in their native lipid context. As our first data show, structural knowledge of protein-lipid interactions for membrane proteins is critical for understanding the structure and function of membrane proteins. This approach has the potential to revolutionize the area of membrane protein research and have a significant impact on membrane protein structure-based medication discovery and development.